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Isolation of Molecular Probes Associated with the Chromosome 15 Instability in the Prader--Willi Syndrome
T. A. Donlon, M. Lalande, A. Wyman, G. Bruns and S. A. Latt
Proceedings of the National Academy of Sciences of the United States of America
Vol. 83, No. 12 (Jun. 15, 1986), pp. 4408-4412
Published by: National Academy of Sciences
Stable URL: http://www.jstor.org/stable/27988
Page Count: 5
You can always find the topics here!Topics: Chromosomes, DNA, DNA probes, Bacteriophages, Cell lines, Libraries, Human chromosomes, In situ hybridization, Genetics, Molecular probes
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Flow cytometry and recombinant DNA techniques have been used to obtain reagents for a molecular analysis of the Prader--Willi syndrome (PWS). HindIII totaldigest libraries were prepared in λ phage Charon 21A from flow-sorted inverted duplicated no. 15 human chromosomes and propagated on recombination-proficient (LE392) and recBC-, sbcB- (DB1257) bacteria. Twelve distinct chromosome 15-specific probes have been isolated. Eight localized to the region 15q11→ 13. Four of these eight sublocalized to band 15q11.2 and are shown to be deleted in DNA of one of two patients examined with the PWS. Heteroduplex analysis of two of these clones, which grew on DB1257 but not on LE392, revealed stem-loop structures in the inserts, indicative of inverted, repeated DNA elements. Such DNA repeats might account for some of the cloning instability of DNA segments from proximal 15q. Analysis of the genetic and physical instability associated with the repeated sequences we have isolated from band 15q11.2 may elucidate the molecular basis for the instability of this chromosomal region in patients with the PWS or other diseases associated with chromosomal abnormalities in the proximal long arm of human chromosome 15.
Proceedings of the National Academy of Sciences of the United States of America © 1986 National Academy of Sciences